Simultaneous expression of cardiac and skeletal muscle isoforms of the L‐type Ca2+ channel in a rat heart muscle cell line.

R. Mejía-Alvarez, Gordon F. Tomaselli, E. Marban

Research output: Contribution to journalArticle

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Abstract

1. We have investigated the identity of the L‐type Ca2+ channels present in the H9c2 myoblast line derived from embryonic rat ventricle. To this end, we characterized macroscopic and unitary Ba2+ currents through Ca2+ channels, and looked for specific genetic messages encoding different L‐type Ca2+ channel isoforms. 2. The macroscopic Ba2+ current (recorded in 10 mM BaCl2) revealed two components with different time courses of activation. The fast component (IBa,fast) activates with a time constant of 23 +/‐ 12 ms (at +10 mV), while the slow component activates with a time constant of 125 +/‐ 12 ms (at +10 mV). 3. Single‐channel recordings revealed the presence of two independent channels with conductance values of 11 and 25 pS (in 70 mM Ba2+). These values are identical to those reported previously for skeletal muscle and cardiac Ca2+ channels, respectively. 4. The mean ensemble current from the 11 pS channel reproduced the time course of the slow component observed at the macroscopic level, while the 25 pS ensemble time course paralleled that of the fast component. 5. Reverse transcriptase polymerase chain reaction (PCR) with alpha 1‐isoform‐specific primers revealed the presence of two distinct transcripts in H9c2 cells. The sequences of the PCR products showed a high degree of homology with the corresponding segments of the rabbit cardiac and skeletal muscle L‐type Ca2+ channel isoforms. Adult rat skeletal and cardiac muscle expressed only one type of transcript. 6. H9c2 cells appear to be unique in that they simultaneously express both skeletal muscle and cardiac isoforms of the L‐type Ca2+ channel alpha 1‐subunit. Thus, the H9c2 cell line may prove to be useful when studying the regulation of subtype‐specific Ca2+ channel gene expression.

Original languageEnglish (US)
Pages (from-to)315-329
Number of pages15
JournalThe Journal of Physiology
Volume478
Issue number2
DOIs
StatePublished - Jul 15 1994
Externally publishedYes

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Cardiac Myocytes
Myocardium
Protein Isoforms
Skeletal Muscle
Cell Line
Myoblasts
Reverse Transcriptase Polymerase Chain Reaction
Rabbits
Gene Expression
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Physiology

Cite this

Simultaneous expression of cardiac and skeletal muscle isoforms of the L‐type Ca2+ channel in a rat heart muscle cell line. / Mejía-Alvarez, R.; Tomaselli, Gordon F.; Marban, E.

In: The Journal of Physiology, Vol. 478, No. 2, 15.07.1994, p. 315-329.

Research output: Contribution to journalArticle

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